Barnyard millet (Echinochloa esculenta) can be processed through soiling, hay, and silage, depending on the weather conditions during harvesting. However, research on barnyard millet is insufficient, and standards for cultivar, seeding density, and fertilizers have not been established. This study was conducted to examine the effects of seeding density and seeding methods on dry matter yields. For this, we used the early-maturing (Shirohie millet) variety of the barnyard millet. The experimental design included different seeding density (10 kg/ha, 15 kg/ha, 20 kg/ha (standard seeding density), 25 kg/ha and 30 kg/ha), and different methods of seeding (drill seeding and broadcast seeding). The seeding date was May 13, 2021, and the harvest date was July 13, 2021. Harvesting was carried out when the heading reached 40 %. Lodging occurred at 5, 9 and 7 at 20, 25 and 30 kg/ha densities in the broadcast seeding, but not in the drill seeding. With decreasing density of seeding, tillage number showed an increasing trend in both drill seeding and broadcast seeding (p>0.05). The plant heights were comparable in both drill seeding and broadcast seeding (p>0.05). The heading stage of barnyard millet was checked July 7 for drill seeding, and, on July 8 for broadcast seeding. It took 62 days, i.e., till July 13 for the heading to reach 40 % of the output. The dry matter yield of barnyard millet was significantly higher at the seeding density of 30 kg/ha, for both the methods of seeding (p<0.05). There was no difference in the chemical composition of grain, based on the seeding method and seeding density. However, as the seeding density increased, the CP, NDF, ADF, and TDN contents increased in both drill seeding and broadcast seeding. We found that, the dry matter content was the highest at the seeding density of 30 kg/ha for both the methods of seeding, but this was only 1.3 times higher than that of 10 kg/ha. Considering the seed price and labor force involved in seeding, it is advisable to have a seeding density of 15-20 kg/ha.

In Sri Lanka, the shoulder in asphalt pavements has been constructed using the materials transported from borrow pit in the iRoad Project due to the low quality of in-situ soils. After excavating 150~200mm thick and 500mm wide shoulder area, the borrow pit materials are placed and compacted according to specifications. The excavated in-situ soils are dumped in designated location. It is estimated that this process of shoulder construction is not economical due to high material transportation cost and can also induce the environmental issues by disposal of in-situ soils. It can also cause distresses such as surface rutting and edge drop-off in soft shoulder section due to bearing capacity failure and off-tracking of vehicle. The heavy rainfall in Sri Lanka can induce severe erosion problem when using the soft shoulder. To improve the strength and durability of pavement shoulders in the iRoad Project, the soil stabilization will be a good alternative to solve the above mentioned problems. The use of in-situ soils with addition of soil stabilizer enables to reduce the construction cost of shoulder section and mitigate the environment issues. The objective of this task is to review the application of soil stabilization method for soft shoulder construction in the iRoad Project. Firstly, the quantitative analysis of soil strength improvement due to soil stabilization was done for soil samples collected from iRoad construction sites. Two types of soils were selected from iRoad Project sites and prepared for soil stabilization testing by the Road Development Authority. Secondly, the appropriate stabilizer was selected at given soil type based on test results. Three different stabilizers, ST-1, ST-2, and ST-3, produced in Korea were used for estimating soil strength improvements. Finally, the optimum stabilizer content was determined for improving shoulder performance. The uniaxial compressive strength (UCS) test was conducted to evaluate the strength of stabilized soil samples in accordance with ASTM D 1633. The use of bottom ash as a stabilizer produced from power plant in Sri Lanka was also reviewed in this task.

The entomopathogenic fungus Metarhizium anisopliae is one of potent biological control agents against a variety ofinsect pests. In this study, we investigated enzyme production of M. anisopliae strains A and B. They produced extracellularenzymes for degrading the epidermis of Monochamus alternatus, a crucial mediator of the pinewood nematode Bursaphelenchusxylophilus. With Q-TOF MS/MS analysis, 29 kDa protein, a major band on a SDS-PAGE gel, was identified as subtilisin-likeserine protease PR1A. M. anisopliae A produced an extracellular enzyme more efficiently than M. anisopliae B: however,enzyme activities targeted for the cuticle were comparable. Our results suggest that the two strains of M. anisopliae havethe biological potential against M. alternatus with insecticidal protease production.

The entomopathogenic fungus Metarhizium anisopliae B is a powerful biological control agent against Monochamusalternatus, a crucial mediator of the pinewood nematode Bursaphelenchus xylophilus. In this study, production of destruxins(dtxs), insecticidal cyclic hexadepsipeptides, was monitored in the submerged culture of M. anisopliae B. Three typesof dtxs, i.e., destruxin A, B, and E, were produced during the culture. Among the three dtxs, the production yield ofdestruxin A was best, followed by destruxin B and E. Destruxin A production was increased when pH was controlledat 6.0, whereas production of destruxin E was not affected by the pH control. The highest yield of dtxs A, B, and Ewere 16.4, 7.3, and 6.1 mg L-1, respectively. Considering that process for dtxs production has not been optimized, M.anisopliae B has more powerful implication as a biocontrol agent.

There are many other detection methods for Bursaphelenchus xylophilus. However, Baermann funnel method, PCR-based methods, which are laborious and time consuming processes that are unsuitable to rapid diagnose the Pine Wilt Disease (PWD) on the field. For these reasons, the aim of our experiment is not only to apply field diagnostic for pine wood nematode (PWN) but also to reduce total time for detection PWN in the pine trees by loop-mediated isothermal amplification (LAMP) with phosphate (Pi)-induced coloration reaction which could be yes or no answer for detection of PWN has not required UV detector but it just could be discriminated by naked eyes within 30 min. Genomic DNA was directly extracted from pine wood chips by Wood Chips Direct Lysis procedure which can be used for LAMP only after simple dilution. Our results suggest that LAMP-Pi detection method, simple and rapid method for detection of PWN, could be applied to the field diagnostic for PWD.

We had evaluated of insecticidal effect from Insecticide-treated net (ITN) that coated by deltamethrin against Monochamus alternatus which serves as a vector for Bursaphelenchus xylophilus. In this experiment, vector has been shown that rapid insecticidal effect response time within 1 hour when they were contacted with ITN. In addition, vectors were showed high mortality within 48 hours. To evaluate insecticidal persistency of ITN and whether releasing insecticide to water, ITNs were soaked in water over various time periods. Water has not shown insecticidal effect to vectors and small amount of insecticide were detected that would not be harmful to other non-target organic being such as honeybee. Also, reduction of insecticidal effect was not observed from the ITN. Taken together, our results suggest that ITN had got highly insecticidal effect to vector as well as could be applied to effectively prevent dispersal of the vector on the field.

Chitin synthase (CHS) is an important enzymatic component, which is required for chitin formation in the cuticles and cuticular linings of other tissues in insects. CHSs have been divided into two classes, class A and B, based on their amino acid sequence similarities and functions. Class A CHS (CHS-A) is specifically expressed in the epidermis and related ectodermal cells such as tracheal cells, while expression of class B CHS (CHS-B) is expressed in gut epithelial cells that produce peritrophic matrices. In this study, we cloned the CHS-A gene from the beet armyworm, Spodoptera exigua. The SeCHS-A mRNA was expressed in all developmental stages and specifically in the epidermis and tracheae tissue by RT-PCR analysis. Expression of SeCHS-A gene was suppressed by feeding double-stranded RNA (dsRNASeCHS-A, 150 ng/larva) in the fifth instar of S. exigua. The suppression of SeCHS-A gene expression significantly induced mortality on pupal stage. Also, larvae fed with dsRNASeCHS-A significantly enhanced pathogenicity of an entomopathogenic fungus, Beauveria bassiana ANU1. These results suggest that the SeCHS-A gene plays an important role in development of S. exigua and dsRNA, which is a specific to SeCHS-A gene, may be applied to effective pest control with B. bassiana.

Oxalic acid has a nematicidal activity against the root-knot nematode Meloidogyne incognita. High producer of oxalic acid was isolated, and then named as Aspergillus niger F22. Oxalic acid production was investigated under various temperatures from 20 – 33oC and rotational speeds in 5 L jar fermenters. Yield of oxalic acid increased with decreasing temperature. The highest yield was obtained at 23oC, showing the yield of oxalic acid of 8.7 g/L, whereas oxalic acid production was least at 33oC. At 20oC, the yield was lower than that of 23oC. At a rotational speed of 300 rpm, serious oxygen depletion was present from 48 - 72 h, resulting in low productivity of 26.2 mg /L·h. When a rotational speed was set at 600 rpm, dissolved oxygen tension was over 40% and oxalic acid production increased up to approximately 55%. Viscosity during the culture differed with temperatures. Viscosity increased with the increment of temperatures. When A. niger F22 was cultured at 23oC, viscosity was 810 cP, which was favorable for oxalic acid production.

It is difficult to identification between Bursaphelenchus spp. and Pine Wood Nematode (PWN) by morphological characteristics without expertise about nematode taxonomy. Furthermore, Baermann funnel method, which is nematode extraction method from wood chips or soil, requires at least 24 hours to extract nematode that is unsuitable to rapid diagnose the Pine Wilt Disease (PWD). For these reasons, the aim of this experiment is not only to improve accuracy of a PCR based method but also to reduce total experiment time for detection Bursaphelenchus spp. and PWN in the wood chips of PWD infected pine tree. In this experiment, we had been employed two PCR primer sets, which were originated from PWN specific Internal Transcribed Spacer (ITS) sequence region and Bursaphenchus spp. universal mitochondrial Cytocrome Oxidase subunit I (mtCOI) sequence region in order to discrimination between Bursaphelenchus spp. and PWN at the same PCR reaction. This experimental procedure was able to reduce experiment time and cost as well as to improve accuracy of detection than previous PCR based detecting method by not using Baermann funnel method and commercial genomic DNA extraction kit but using direct pine wood chips lysis method.